Investigation of the mechanism of fangfeng tongsheng pills in the treatment COVID-19 by network pharmacology and molecular docking

Objective: To explore the active components and potential mechanism of Fangfeng Tongsheng Pills by using network pharmacology and molecular docking in the treatment of coronavirus disease 19(COVID-19). Methods The main chemical constituents and action targets of various medicines in Fangfeng Tongsheng Pills were collected via traditional Chinese medicine system pharmacology database and online analysis platform(TCMSP). The related targets of COVID-19 were collected by using GeneCards database, and the repeating parts with Fangfeng Tongsheng Pills were taken as the research targets. Cytoscape software was used to create a drug-target-disease network. The common target was imported into STRING database, and the protein-protein interaction network diagram was constructed by Cytoscape software. The GO(gene ontology) function enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were performed by DAVID to predict their mechanism. The core components of Fangfeng Tongsheng Pills were docked with the therapeutic target of COVID-19 by AutoDock software. Results A total of 224 active compounds and 696 active targets were screened from Fangfeng Tongsheng Pills, including 79 targets coincided with COVID-19, and 10 active compounds, i.e. quercetin, luteolin, kaempferol,beta-sitosterol, naringenin, etc., 23 effective targets, i.e. PTGS2, PTGS1, NOS2, F10, DPP4, etc. A total of 65 GO function enrichment analysis results and 101 KEGG pathway enrichment results were obtained, including inflammatory response, tumor necrosis factor(TNF) signaling pathway, hypoxia inducible factor-1(HIF-1) signaling pathway, vascular endothelial growth factors(VEGF) signaling pathway, toll-like receptors(TLRs) signaling pathway, phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt) signaling pathway, and mitogen-activated protein kinase(MAPK) signaling pathway. Conclusion The active components in Fangfeng Tongsheng Pills, such as beta-sitosterol, quercetin, luteolin, kaempferol and naringenin, can combine with SARS-Co V2-3CL hydrolase and ACE2, act on the key target [TNF, Caspase-3, mitogen-activated protein kinase(MAPK1), interleukin-6(IL-6), prostaglandin-endoperoxide synthase 2(PGTS2)] of TNF, HIF-1, VEGF, MAPK and toll-like receptor signaling pathway, and play the roles of anti-inflammation, immune regulation, anti-hypoxic stress and anti-virus infection, thus play a role in the treatment of COVID-19.

Sex-based clinical and immunological differences in COVID-19.

BACKGROUND: Males and females differ in their immunological responses to foreign pathogens. However, most of the current COVID-19 clinical practices and trials do not take the sex factor into consideration. METHODS: We performed a sex-based comparative analysis for the clinical outcomes, peripheral immune cells, and severe acute respiratory syndrome coronavirus (SARS-CoV-2) specific antibody levels of 1558 males and 1499 females COVID-19 patients from a single center. The lymphocyte subgroups were measured by Flow cytometry. The total antibody, Spike protein (S)-, receptor binding domain (RBD)-, and nucleoprotein (N)- specific IgM and IgG levels were measured by chemiluminescence. RESULTS: We found that male patients had approximately two-fold rates of ICU admission (4.7% vs. 2.7% in males and females, respectively, P = 0.005) and mortality (3% vs. 1.4%, in males and females, respectively, P = 0.004) than female patients. Survival analysis revealed that the male sex is an independent risk factor for death from COVID-19 (adjusted hazard ratio [HR] = 2.22, 95% confidence interval [CI]: 1.3-3.6, P = 0.003). The level of inflammatory cytokines in peripheral blood was higher in males during hospitalization. The renal (102/1588 [6.5%] vs. 63/1499 [4.2%], in males and females, respectively, P = 0.002) and hepatic abnormality (650/1588 [40.9%] vs. 475/1499 [31.7%], P = 0.003) were more common in male patients than in female patients. By analyzing dynamic changes of lymphocyte subsets after symptom onset, we found that the percentage of CD19+ B cells and CD4+ T cells was generally higher in female patients during the disease course of COVID-19. Notably, the protective RBD-specific IgG against SARS-CoV-2 sharply increased and reached a peak in the fourth week after symptom onset in female patients, while gradually increased and reached a peak in the seventh week after symptom onset in male patients. CONCLUSIONS: Males had an unfavorable prognosis, higher inflammation, a lower percentage of lymphocytes, and indolent antibody responses during SARS-CoV-2 infection and recovery. Early medical intervention and close monitoring are important, especially for male COVID-19 patients.

Implications of cardiac markers in risk-stratification and management for COVID-19 patients.

BACKGROUND: COVID-19 has resulted in high mortality worldwide. Information regarding cardiac markers for precise risk-stratification is limited. We aim to discover sensitive and reliable early-warning biomarkers for optimizing management and improving the prognosis of COVID-19 patients. METHODS: A total of 2954 consecutive COVID-19 patients who were receiving treatment from the Wuhan Huoshenshan Hospital in China from February 4 to April 10 were included in this retrospective cohort. Serum levels of cardiac markers were collected after admission. Coronary artery disease diagnosis and survival status were recorded. Single-cell RNA-sequencing and bulk RNA-sequencing from different cohorts of non-COVID-19 were performed to analyze SARS-CoV-2 receptor expression. RESULTS: Among 2954 COVID-19 patients in the analysis, the median age was 60 years (50-68 years), 1461 (49.5%) were female, and 1515 (51.3%) were severe/critical. Compared to mild/moderate (1439, 48.7%) patients, severe/critical patients showed significantly higher levels of cardiac markers within the first week after admission. In severe/critical COVID-19 patients, those with abnormal serum levels of BNP (42 [24.6%] vs 7 [1.1%]), hs-TNI (38 [48.1%] vs 6 [1.0%]), α- HBDH (55 [10.4%] vs 2 [0.2%]), CK-MB (45 [36.3%] vs 12 [0.9%]), and LDH (56 [12.5%] vs 1 [0.1%]) had a significantly higher mortality rate compared to patients with normal levels. The same trend was observed in the ICU admission rate. Severe/critical COVID-19 patients with pre-existing coronary artery disease (165/1,155 [10.9%]) had more cases of BNP (52 [46.5%] vs 119 [16.5%]), hs-TNI (24 [26.7%] vs 9.6 [%], α- HBDH (86 [55.5%] vs 443 [34.4%]), CK-MB (27 [17.4%] vs 97 [7.5%]), and LDH (65 [41.9%] vs 382 [29.7%]), when compared with those without coronary artery disease. There was enhanced SARS-CoV-2 receptor expression in coronary artery disease compared with healthy controls. From regression analysis, patients with five elevated cardiac markers were at a higher risk of death (hazards ratio 3.4 [95% CI 2.4-4.8]). CONCLUSIONS: COVID-19 patients with pre-existing coronary artery disease represented a higher abnormal percentage of cardiac markers, accompanied by high mortality and ICU admission rate. BNP together with hs-TNI, α- HBDH, CK-MB and LDH act as a prognostic biomarker in COVID-19 patients with or without pre-existing coronary artery disease.